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Research > Research Cores > Flow Cytometry and Fluorescence Activated Cell Sorting > Cell Sorting

Tips for Successful Sorting

  • Book your sort time early! 7-10 days in advance will ensure you will get your desired day and time.
  • You must cancel 24 hours in advance or you will be billed for the time you have reserved.
  • Bring a negative control, as well as positive controls for multi-color sorts for compensation. If you feel confident that you can choose the cells you want without these controls, you may do so.
  • If you wish to bring your cells in the tubes for the sorter, filter top tubes that are compatible with the VantageSE are the following: Falcon catalog #2235 (I order them from Fisher; catalog number 08-771-23, 1 case of 500 is $338.60).
  • Filter your cells and keep them cold to keep them from clumping, thus resulting in better yield and purity.
Tips to reduce cell clumping

Wash cells to be sorted to be sorted with cold serum free calcium and magnesium free HBSS/high glucose. Resuspend in the same cold wash buffer with 0.1mM EDTA and less than 2% serum.

Prior to staining

Collagenase type II (Sigma)70 mg/25ml of serum free calcium and magnesium free HBSS/high glucose can be used to gently pry apart aggregated cells. The cells should be incubated at 37OC with gentle mixing every five minutes for a total of fifteen minutes. The cells should be washed as described above and stained at 4OC and kept cold.

For Sorting, You Will Need to Bring the Following

  • Negative control (at least 0.5ml, if possible)
  • Positive controls for each fluorochrome you are using
  • Ice
  • Media of your choice to collect into (adding antibiotics is recommended)
For regular speed sorting, samples should contain less than 2% FBS at 5 - 10 million cells/mL

For high speed sorting, samples should contain less than 2% FBS at 15 - 20 million cells/mL



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